Introduction:

Several physiological phenomena in the body can be described using electrical models. In 1952,
Hodgkin and Huxley were able to accurately predict the depolarization of neurons based on the flux of
different ions across the cell membrane. These depolarizations, as well as the propagation of signals
throughout the nervous system, were shown to be dependent on the ionic gradient between the intra- and
extra-cellular environment. In this experiment, after familiarizing ourselves with basic components of
circuits, we constructed a voltage clamp circuit capable of generating a current through a fluid chamber
across an unknown impedance. After confirming the functionality of our circuit, we used it to measure
the impedances across both a section of Teflon film and a lipid bilayer. The application of the voltage
clamp in these settings confirmed Hodgkin and Huxley’s findings and demonstrated membrane
depolarization at a macroscopic level.

Methods:

Basic Circuits

Before preparing a voltage clamp circuit, we first reviewed basic circuit elements and verified
several foundations of electronics, such as Ohm’s Law and Kirchoff’s Current and Voltage Laws. To do
this, a voltage divider circuit was built with 330 Ω and 470 Ω resistors and analyzed for both an AC and
DC input. We then constructed an RC circuit with a 330 Ω resistor and 1 nF capacitor and measured the
impedance and voltage across both the capacitor and the resistor for a variety of frequencies with 9 V
(peak-to-peak) AC and DC inputs of various waveforms. Finally, inverting and non-inverting amplifiers,
were tested and their ability to achieve a voltage gain confirmed for various resistances (1 k Ω and 2 kΩ)
and input voltages.

Using the Voltage Clamp for a Known and Unknown Impedance

After constructing the voltage clamp, we generated a current across a fluid chamber using a V ref of
0.5 V and an Rref of 1 MΩ. A 390 Ω resistor was used as Rclamp and vout was observed and recorded for a
triangular input waveform of constant amplitude and period. The resistance of this clamp was then
measured and compared to the theoretical value of R clamp. The capacitance of a 1 nF capacitor Cclamp was
also measured and compared to the labeled value following a similar procedure.

After washing the components of the fluid chamber, we used our voltage clamp and Ag-AgCl
electrodes and a 1 M Ω Rclamp, to observe the role of Teflon film as a capacitor in low-level 1 M NaCl
solution. Given a 200 mV peak-to-peak vref and knowing the area of the Teflon film and the distance
between the solutions, it was possible to find the capacitance and impedance of the film.

Using the Voltage Clamp to finding the Impedance Across a Lipid Bilayer

In a similar experimental setup, we used a combination of phosphatidyl choline (PC) and

cholesterol as lipids and primed the perforation in the Teflon film with a 1:10 solution of
hexadecane:pentane. Using the oscilloscope, we were able to confirm that the hydrophobic lipids formed
a bilayer on the perforation to avoid any interactions with our aqueous solution. In addition to testing
various DC and AC voltages across this bilayer to simulate membrane depolarization, we found the area
of the perforation in order to calculate C/A for the bilayer.